CN115919951B - Traditional Chinese medicine composition and preparation method and application thereof - Google Patents
Traditional Chinese medicine composition and preparation method and application thereof Download PDFInfo
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- CN115919951B CN115919951B CN202211519371.1A CN202211519371A CN115919951B CN 115919951 B CN115919951 B CN 115919951B CN 202211519371 A CN202211519371 A CN 202211519371A CN 115919951 B CN115919951 B CN 115919951B
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Classifications
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Medicines Containing Plant Substances (AREA)
Abstract
The invention provides a traditional Chinese medicine composition, and a preparation method and application thereof. The traditional Chinese medicine composition formula provided by the invention comprises the following raw materials in parts by weight: 10-15 parts of beautyberry, 8-15 parts of coptis chinensis, 12-20 parts of radix scutellariae, 10-18 parts of pericarpium granati, 10-20 parts of garden burnet and 8-12 parts of liquorice, and the traditional Chinese medicine composition has the effects of clearing heat and detoxicating, and also has the effects of astringing intestines to stop diarrhea, cooling blood and stopping bleeding, so that recovery of the organism in the clinical pathogenic escherichia coli infection course is accelerated; in addition, the invention can promote the release of the active ingredients of the traditional Chinese medicine to a greater extent through ultrasonic-assisted composite enzymolysis extraction, strengthen the antibacterial activity of the traditional Chinese medicine, more effectively treat the yellow-white dysentery of pigs caused by bacterial infection and play a more remarkable clinical prevention and treatment effect.
Description
Technical Field
The invention belongs to the technical field of veterinary medicines, and particularly relates to a traditional Chinese medicine composition, and a preparation method and application thereof.
Background
Diarrhea of piglets is one of the most common diseases in the global pig industry, and the diarrhea of piglets can be caused by infection of escherichia coli, salmonella, porcine epidemic diarrhea virus, transmissible gastroenteritis virus, coccidia and the like. The enterotoxigenic escherichia coli (enterotoxigenic Escherichia coli, ETEC) is always considered as one of important pathogens causing diarrhea of piglets, and after the ETEC invades the intestinal tract of the piglets, the enterotoxigenic escherichia coli can be planted in the intestinal tract through pili, so that enterotoxigenic escherichia coli can be secreted to cause electrolyte imbalance of intestinal epithelial cells, and finally, the piglets can have clinical symptoms represented by yellow diarrhea and white diarrhea. Yellow scour of piglets mainly occurs in 1-7 days old, usually the whole litter is ill, and the mortality rate is up to 90%; white diarrhea of piglets is mostly caused by 10-30 days old, the morbidity is over 80 percent, the mortality is lower, but the white diarrhea can still have a certain influence on the growth performance of the piglets, and the white diarrhea can reduce important indexes such as daily weight gain of the piglets, weaning weight of the piglets and the like.
At present, the escherichia coli infection is mainly treated clinically through vaccination prevention and antibiotics, but the vaccine is often not efficient due to the fact that the escherichia coli serotypes are numerous, and the escherichia coli drug resistance phenomenon and the drug residue problem caused by the overuse of antibiotics are threatened to public health safety and human health. Therefore, development of the antibiotic-replacing medicament for effectively controlling the infection of the escherichia coli drug-resistant strain and reducing the bacterial drug resistance has wide clinical application prospect.
The Chinese herbal medicine has long application history in China, is recorded in Chinese pharmacopoeia, has the functions of clearing heat and detoxicating, cooling blood and stopping bleeding, astringing intestines and stopping diarrhea, tonifying spleen and replenishing qi, astringing lung and relieving asthma, enriching blood and nourishing blood and the like, and has the advantages of safety, richness, low cost and difficult drug resistance due to the fact that the Chinese herbal medicine is derived from natural plants, and has obvious clinical treatment effects on clinical diseases through differentiation of Chinese medicine theory and further on evidence medication. Yellow and white scour of piglets is mostly caused by warm epidemic toxin hidden in sow bodies, and primary diseases and secondary diseases or primary piglets Wei Wai are weakened in capability due to cold weather, narrow houses, dark and moist pollution, and are infected by external damp-heat toxin. Damp heat is accumulated in the large intestine, and damages the intestinal tract to cause dysfunction, and diarrhea and dysentery occur due to clear turbidity. The disease is sensitive to damp-heat toxin, so that the disease is characterized by stink and sticky and yellow or white stool. Heat is a yin fluid easily damaged by pathogenic yang, so that symptoms of yin deficiency such as thirst, pi Ganmao dryness and the like quickly appear, and coma and death are caused by yin deficiency to the utmost extent and yin-yang separation. According to the second part of the 'traditional Chinese veterinary drug classical 2020 edition' recorded dysentery stopping powder, the formula comprises 40g of realgar, 110g of wrinkled giant hyssop, 150g of talcum and 100g of Chinese gentian root, wherein the formula also comprises 600g of pulsatilla root, 300g of gentian root and 100g of coptis root, and the two classical formulas have the effects of clearing heat and detoxicating and resolving dampness and stopping dysentery. Compared with powder, the Chinese pulsatilla root oral liquid in pharmacopoeia has better effect of treating yellow and white dysentery by drenching piglets, the formula comprises 300g of Chinese pulsatilla root, 150g of coptis chinensis, 300g of ash bark and 225g of amur corktree bark, has the effects of clearing heat and detoxicating, cooling blood and stopping dysentery, has higher cost in clinical application, ensures that the application popularity of the Chinese medicinal preparation is not high, has relatively low price, but causes single medicinal composition types, and has unstable treatment effect when the clinical application faces pathogenic change.
Through patent search, some patent documents are found to disclose traditional Chinese medicine formulas with potential treatment or prevention effects on livestock diarrhea. For example, a traditional Chinese medicine for treating pullorum disease has been disclosed in the prior patent, which comprises: 1kg of coptis chinensis, 1kg of phellodendron, 1kg of scutellaria baicalensis, 1kg of common andrographis herb, bai Weng kg of 2kg of bighead atractylodes rhizome, 2kg of white paeony root, 2kg of pericarpium granati, 2kg of dandelion, 2kg of liquorice, 2kg of garden burnet, 0.5kg of magnolia officinalis, 0.5kg of red halloysite, 0.5kg of fructus kochiae, 2.5kg of radix isatidis, 2.5kg of dyers woad leaf and 2.5kg of dried orange peel, the traditional Chinese medicine composition has an obvious cure effect on pullorum disease and has no adverse reaction; the prior patent discloses a pure traditional Chinese medicine formula for treating various animal diarrhea and a preparation method thereof, comprising the following steps: 1g of white peony root, 1g of bighead atractylodes rhizome, 1g of pulsatilla root, 1g of rhizoma atractylodis, 2g of dried orange peel, 1g of dyers woad leaf, 1g of amur corktree bark, 0.75g of weeping forsythiae capsule, 3g of dandelion, 1.5g of ash bark, 5g of garden burnet, 1g of pomegranate rind and 1g of poria cocos, wherein the traditional Chinese medicine compound has obvious curative effects on various diarrhea diseases of animals, such as yellow-white diarrhea of piglets, epidemic diarrhea, transmissible gastroenteritis and the like; also disclosed is a method for preparing a Chinese medicinal oral liquid for treating diarrhea of piglets, comprising the following steps: 350g of polygonum, 200g of valeriana jatamansi, 200g of oleaster leaves, 150g of bistort rhizome and 100g of cactus, and the traditional Chinese medicine compound oral liquid reduces diarrhea disease mortality of piglets and improves the breeding benefit of live pigs.
Through analysis of the prior art, the traditional Chinese medicine composition for preventing and treating piglet yellow-white dysentery in the disclosed patent is more, generally consists of more than 10 traditional Chinese medicines, and has a certain clinical effect, but the traditional Chinese medicines with excessive tastes cause cost increase, so that the application at the breeding end and the huge requirement for matching the breeding end are difficult to realize. Aiming at the problems, the liquid traditional Chinese medicine composition with high cost performance and stable curative effect is urgently required to be developed, and the preparation method is optimized, so that the effective prevention and treatment of the yellow-white dysentery of piglets are realized, the cost of traditional Chinese medicines can be controlled, the requirements of actual breeding production work are met, and the liquid traditional Chinese medicine composition has important significance in promoting the healthy development of the pig industry.
Disclosure of Invention
The present invention aims to solve at least one of the technical problems in the prior art described above. Therefore, the first aspect of the invention provides a traditional Chinese medicine composition which can be used for preventing and treating yellow and white scour of piglets, has obvious prevention effect in clinical application, quick treatment effect and high cure rate, and has no obvious side effect on production.
The second aspect of the invention provides a preparation method of the traditional Chinese medicine composition.
The third aspect of the invention provides a medicament for preventing and treating swine yellow-white dysentery.
The fourth aspect of the invention provides application of the traditional Chinese medicine composition.
According to a first aspect of the present invention, a traditional Chinese medicine composition is provided, wherein the formula of the traditional Chinese medicine composition comprises the following raw materials in parts by weight: 10-15 parts of beautyberry, 8-15 parts of coptis chinensis, 12-20 parts of radix scutellariae, 10-18 parts of pericarpium granati, 10-20 parts of garden burnet and 8-12 parts of liquorice.
In some embodiments of the present invention, the formulation of the traditional Chinese medicine composition is composed of the following raw materials in parts by weight: 11-13 parts of beautyberry, 9-11 parts of coptis chinensis, 13-15 parts of baikal skullcap root, 12-16 parts of pericarpium granati, 14-16 parts of garden burnet and 9-11 parts of liquorice.
In the invention, beautyberry is selected, and the beautyberry has the effects of detoxification, inflammation diminishing, astringency and hemostasis, and is mainly used for treating gastrointestinal bleeding, traumatic bleeding and other symptoms; then selecting coptis chinensis, having bitter and cold property, having the effects of clearing heat and drying dampness, purging fire and detoxifying, and mainly treating damp-heat diarrhea, heart fire exuberance, stomach fire exuberance, liver and gall damp-heat, conjunctival congestion and swelling and pain, fire toxin sore and carbuncle and other symptoms; radix Scutellariae is selected, which is bitter and cold in nature, has the effects of clearing heat and drying dampness, purging pathogenic fire and removing toxin, and stopping bleeding, and is mainly used for treating symptoms such as gastrointestinal damp-heat, cough due to lung heat, high fever and greedy drink; the pomegranate rind is selected, is sour, astringent and warm, has the effects of relieving diarrhea with astringents, stopping bleeding and expelling parasites, and is mainly used for treating diarrhea and hematochezia; in addition, the garden burnet is bitter, sour, astringent and slightly cold in nature, has the effects of cooling blood, removing toxicity, stopping bleeding and astringing wound, and is mainly used for treating symptoms such as bloody dysentery, scalds and the like; and licorice is selected, so that the Chinese medicinal preparation is sweet in nature, has the effects of tonifying spleen and replenishing qi, harmonizing middle energizer and relieving urgency and detoxifying, and is mainly used for treating symptoms such as weakness of spleen and stomach, poisoning and the like.
In the recipe, coptis root and baikal skullcap root are used as monarch drugs and have the effects of clearing heat and drying dampness and purging pathogenic fire and removing toxin; the garden burnet and the callicarpa nudiflora are ministerial drugs, and have the effects of cooling blood, removing toxicity, stopping bleeding and astringing; the pericarpium Granati is an adjuvant drug with the effects of astringing intestine to stop diarrhea and stopping bleeding, and the licorice is a guiding drug for invigorating spleen and replenishing qi and harmonizing various drugs. The medicines are combined together to play roles of clearing heat and detoxicating, astringing intestine and stopping diarrhea, cooling blood and stopping bleeding, and accelerating organism recovery.
According to a second aspect of the present invention, there is provided a method for preparing the Chinese medicinal composition according to the first aspect, comprising:
Taking the powder of beautyberry, baical skullcap root, golden thread, pomegranate rind, garden burnet root and liquoric root according to the proportion,
S1: mixing powder of Callicarpa nudiflora, coptidis rhizoma, pericarpium Granati, radix Sangusorbae and Glycyrrhrizae radix, adding water, ultrasonic extracting, and performing enzymolysis with complex enzyme to obtain extractive solution A and residue B;
S2: extracting Scutellariae radix powder with water to obtain extractive solution C and residue D;
s3: mixing the residue B and the residue D, extracting with water to obtain extractive solution E;
S4: mixing the extract A, the extract C and the extract E, and concentrating to obtain the traditional Chinese medicine composition.
In some embodiments of the invention, the fineness of the powder is 30 mesh to 40 mesh.
In some embodiments of the present invention, the ratio of the volume of the added water in S1 to the total mass of the coptis chinensis, pericarpium granati, sanguisorba officinalis and licorice powder is 8mL to 12mL:1g.
In some embodiments of the invention, the temperature of the ultrasonic extraction in S1 is 50 ℃ to 60 ℃.
In some embodiments of the invention, the power of the ultrasonic extraction in S1 is 100W to 180W.
In some embodiments of the invention, the time of the ultrasonic extraction in S1 is 30min to 40min.
In some embodiments of the invention, the complex enzyme in S1 comprises at least three enzymes of cellulase, pectinase, amylase, glycosidase, mannanase, papain.
In some embodiments of the invention, the enzyme activity of each enzyme in the complex enzyme in S1 is independently 9 to 15 kiloU/g.
In some embodiments of the invention, the amount of the complex enzyme added in S1 is 3% to 5% of the mass of the extract a.
In some embodiments of the invention, in the water extraction of S2, the ratio of the volume of water to the mass of the baikal skullcap root powder is 8mL to 12mL: 1g) .
In some embodiments of the invention, the water extraction time of S2 is 1.5h to 2h.
In some preferred embodiments of the invention, the complex enzyme in S1 is cellulase, pectinase, papain.
In some preferred embodiments of the present invention, when the complex enzyme in S1 is cellulase, pectinase, papain, the mass ratio of the three is (1-3): (1-5): (1-5).
In some more preferred embodiments of the present invention, when the complex enzyme in S1 is cellulase, pectinase, papain, the mass ratio of the three is (1-2): (1-2): (3-4).
In some more preferred embodiments of the invention, the conditions for the enzymatic hydrolysis of the complex enzyme in S1 are: the pH value is 4.5-5.0, the temperature is 50-60 ℃ and the time is 1-3 h.
In some more preferred embodiments of the present invention, in the mixed aqueous extraction of S3, the ratio of the volume of water to the total mass of the residues B and D is 6mL to 8mL:1g.
In some more preferred embodiments of the present invention, the time for the mixed water extraction of S3 is 1h to 1.5h.
In some more preferred embodiments of the present invention, the total mass ratio of the volume of the concentrated Chinese medicinal composition obtained in S3 to the added powders of scutellaria baicalensis, coptis chinensis, pericarpium granati, sanguisorba officinalis and liquorice is (1-2) mL:1g.
According to a third aspect of the present invention, there is provided a medicament for preventing and treating swine yellow-white dysentery, wherein the active ingredients in the medicament comprise the Chinese medicinal composition of the first aspect.
In some embodiments of the invention, the medicament further comprises a pharmaceutically acceptable excipient.
In some embodiments of the invention, the pharmaceutically acceptable excipients include, but are not limited to, diluents, binders, disintegrants, lubricants, excipients, fillers.
In some embodiments of the invention, the dosage form of the medicament is selected from one of a powder, a tablet, an aerosol, a mixture, and a granule.
According to a fourth aspect of the present invention, there is provided the use of a Chinese medicinal composition according to the first aspect in the manufacture of a medicament for the prevention and/or treatment of at least one of (a) to (c):
(a) Yellow dysentery of pigs;
(b) White diarrhea of pigs;
(c) Bacterial infection in pigs.
In some embodiments of the invention, the bacteria in (c) are selected from at least one of escherichia coli, staphylococcus aureus, salmonella.
In some preferred embodiments of the invention, the pig is a piglet.
The beneficial effects of the invention are as follows:
the traditional Chinese medicine composition has the effects of clearing heat and detoxicating, astringing intestines to check diarrhea, cooling blood and stopping bleeding, has obvious prevention and treatment effects on piglet yellow-white dysentery, and enhances piglet immunity; the invention can promote the release of the active ingredients of the traditional Chinese medicine to a greater extent by adopting the ultrasonic-assisted compound enzymolysis liquid extraction method, and can exert more remarkable clinical prevention and treatment effects compared with the traditional water extraction method.
Drawings
The invention is further described with reference to the accompanying drawings and examples, in which:
FIG. 1 shows the characteristics of yellow dysentery feces of 1-7-day-old piglets.
FIG. 2 shows the diarrhea cure criteria for piglets of 1-7 days of age.
FIG. 3 shows baicalin chromatograms of samples 1 to 7 in the test examples of the present invention.
FIG. 4 shows the chromatograms of berberine hydrochloride of samples 1 to 7 in the test example of the present invention.
Detailed Description
The conception and the technical effects produced by the present invention will be clearly and completely described in conjunction with the embodiments below to fully understand the objects, features and effects of the present invention. It is apparent that the described embodiments are only some embodiments of the present invention, but not all embodiments, and that other embodiments obtained by those skilled in the art without inventive effort are within the scope of the present invention based on the embodiments of the present invention.
Example 1
The embodiment prepares a traditional Chinese medicine composition, and the formula comprises the following components in parts by weight: 12 parts of beautyberry, 10 parts of coptis chinensis, 15 parts of radix scutellariae, 15 parts of pericarpium granati, 15 parts of sanguisorba officinalis and 10 parts of liquorice.
The specific process is as follows:
Crushing medicinal materials: the beautyberry, coptis chinensis, scutellaria baicalensis, pericarpium granati, sanguisorba officinalis, liquorice and the like are subjected to pretreatment such as washing, cutting, drying and the like according to the traditional Chinese herbal medicine processing technology, and then the medicinal materials are respectively crushed and sieved by a 30-mesh sieve.
And (3) ultrasonic treatment: sequentially weighing the medicinal materials according to the proportion of the formula (except for scutellaria baicalensis, which is required to be extracted singly), stirring and mixing uniformly, adding water with the mass of 10 times of the volume of the medicinal materials, performing ultrasonic extraction at the temperature of 55 ℃, controlling the ultrasonic power to be 150W and the ultrasonic time to be 40min to obtain an extract containing medicinal residues, adjusting the pH value to 4.5-5.0, adding compound enzyme (cellulase (10 ten thousand U/g) with the mass of 4% of the total mass of the medicinal materials, pectase (9 ten thousand U/g) and xylose protease (10 ten thousand U/g) with the mass ratio of = 1:1.5:3), starting an enzymolysis procedure, stirring in an enzymolysis tank while performing enzymolysis for 3h, controlling the enzymolysis temperature to be 55+/-3 ℃, and performing solid-liquid separation to collect the extract A and the medicinal residues B after enzymolysis is completed;
Extracting radix Scutellariae alone: adding water with the volume 10 times of that of radix Scutellariae in the formula, heating to slight boiling, adding radix Scutellariae, slightly boiling for 1 hr, collecting extractive solution C, and storing radix Scutellariae residue D;
adding distilled water with volume 10 times of that of the residue B, heating to slight boiling, adding Scutellariae radix residue D, extracting for 1 hr, and collecting extractive solution E.
Mixing the extract A, C and E, concentrating at normal temperature according to the mass-volume ratio of 1g to 2mL of the total mass and the concentrated volume of the medicinal materials to obtain the traditional Chinese medicine composition (the concentration is 0.5 g/mL), adding 0.3% sodium benzoate and 4% tween-80, sterilizing and packaging to obtain the finished product.
Example 2
The embodiment prepares a traditional Chinese medicine composition, and the formula comprises the following components in parts by weight: 15 parts of beautyberry, 15 parts of coptis chinensis, 15 parts of radix scutellariae, 10 parts of pericarpium granati, 10 parts of sanguisorba officinalis and 10 parts of liquorice. The specific procedure was carried out with reference to example 1.
Example 3
The embodiment prepares a traditional Chinese medicine composition, and the formula comprises the following components in parts by weight: 12 parts of beautyberry, 10 parts of coptis chinensis, 15 parts of radix scutellariae, 15 parts of pericarpium granati, 15 parts of sanguisorba officinalis and 10 parts of liquorice. Cellulase: pectase: the mass ratio of the xylose protease is 1:2:2. The specific procedure was carried out with reference to example 1.
Comparative example 1
The comparative example prepared a traditional Chinese medicine composition, which is different from example 1 in that the formula is as follows in parts by weight: 5 parts of beautyberry, 15 parts of coptis chinensis, 10 parts of radix scutellariae, 20 parts of pericarpium granati, 15 parts of garden burnet and 10 parts of liquorice. Other references are to example 1.
Comparative example 2
The comparative example prepared a traditional Chinese medicine composition, which is different from example 1 in that the formula is as follows in parts by weight: 10 parts of beautyberry, 5 parts of coptis chinensis, 15 parts of radix scutellariae, 12 parts of pericarpium granati, 20 parts of garden burnet and 10 parts of liquorice. Other references are to example 1.
Comparative example 3
The comparative example prepared a traditional Chinese medicine composition, which is different from example 1 in that the formula is as follows in parts by weight: 10 parts of coptis chinensis, 15 parts of scutellaria baicalensis, 15 parts of pericarpium Granati, 15 parts of sanguisorba officinalis and 10 parts of liquorice. Other references are to example 1.
Comparative example 4
The comparative example prepared a traditional Chinese medicine composition, which is different from example 1 in that the pericarpium Granati is replaced by gallnut which is equal in weight part and contains a large amount of tannic acid, and the formula is as follows: 12 parts of beautyberry, 10 parts of coptis chinensis, 15 parts of radix scutellariae, 15 parts of gallnut, 15 parts of sanguisorba officinalis and 10 parts of liquorice. Other references are to example 1.
Comparative example 5
The comparative example prepared a traditional Chinese medicine composition, which is different from example 1 in that the sanguisorba is replaced by gallnut which is equal in weight part and contains a large amount of tannic acid, and the formula is as follows: 12 parts of beautyberry, 10 parts of coptis chinensis, 15 parts of baikal skullcap root, 15 parts of pericarpium Granati, 15 parts of Chinese gall and 10 parts of liquorice. Other references are to example 1.
Comparative example 6
The comparative example prepares a traditional Chinese medicine composition, which is different from the example 1 in that 10 parts of honeysuckle and 15 parts of fructus forsythiae are added to replace coptis and baikal skullcap root respectively, and the formula is as follows: 12 parts of beautyberry, 10 parts of honeysuckle, 15 parts of weeping forsythiae capsule, 15 parts of pomegranate rind, 15 parts of garden burnet and 10 parts of liquorice. Other references are to example 1.
Comparative example 7
The comparative example prepared a traditional Chinese medicine composition, which is different from example 1 in that 30 parts of pulsatilla root is added to replace baikal skullcap root and garden burnet root, and the formula is: 12 parts of beautyberry, 10 parts of coptis chinensis, 30 parts of pulsatilla chinensis, 15 parts of pericarpium granati and 10 parts of liquorice.
The specific process is as follows:
crushing medicinal materials: the beautyberry, coptis chinensis, pulsatilla chinensis, pericarpium granati, liquorice are subjected to pretreatment such as washing, cutting, drying and the like according to the traditional Chinese herbal medicine processing technology, and then the medicinal materials are respectively crushed and sieved by a 30-mesh sieve.
And (3) ultrasonic treatment: sequentially weighing the medicinal materials according to the formula ratio, stirring and mixing uniformly, adding water with the mass of 10 times of the volume of the medicinal materials, performing ultrasonic extraction at the temperature of 55 ℃ and ultrasonic power of 150W for 40min to obtain an extract containing medicinal residues, regulating the pH to 4.5-5.0, adding compound enzyme (cellulase (10 ten thousand U/g) of which the mass is 4% of the total mass of the medicinal materials (pectase (9 ten thousand U/g) of which the mass ratio is 10 ten thousand U/g) of pectase (10 ten thousand U/g) of which the mass ratio is 1:1.5:3), starting an enzymolysis procedure, performing enzymolysis for 3h in an enzymolysis tank while stirring, controlling the enzymolysis temperature to 55+/-3 ℃, and performing solid-liquid separation to collect the extract A and the medicinal residues B after enzymolysis is completed;
adding distilled water 10 times of the volume of the residue B, heating to slight boiling, extracting for 1 hr, and collecting extractive solution C.
Mixing the extract A and the extract C, concentrating at normal temperature according to the mass-volume ratio of 1g to 2mL of the total mass and the concentrated volume of the medicinal materials in the formula to obtain the traditional Chinese medicine composition (the concentration is 0.5 g/mL), adding 0.3% sodium benzoate and 4% tween-80, sterilizing and packaging to obtain the finished product.
Comparative example 8
The comparative example prepared a traditional Chinese medicine composition, which is different from example 1 in that hemicellulase was used instead of cellulase, and in the complex enzyme, the hemicellulase: pectase: the amylase mass ratio is 1:1.5:3.
Comparative example 9
The comparative example prepared a traditional Chinese medicine composition, which is different from example 1 in that the cellulase in the complex enzyme: pectase: the mass ratio of the xylose protease is 1:3:6.
Test examples
1. In vitro bacteriostatic Activity evaluation
The minimum inhibitory concentration MIC and minimum bactericidal concentration MBC of the traditional Chinese medicine compositions prepared in examples 1 to 3 and comparative examples 1 to 9 on staphylococcus aureus (CICC 10384), salmonella quality control strain (CICC 10437), escherichia coli quality control strain (ATCC 25922), escherichia coli K88 strain were measured by a broth dilution method, 100 mu L of MH broth culture medium was added to columns 2 to 11 of a 96-well plate, 200 mu L of MH broth culture medium was added to column 12 as a control, 500mg/mL of 100 mu L of traditional Chinese medicine composition liquid was added to column 1, and the final concentration of the traditional Chinese medicine composition liquid in columns 1 to 11 was 250, 125, 62.50, 31.25, 15.63, 7.82, 3.91, 1.96, 0.98, 0.49 and 0mg/mL, respectively, and the final concentration of bacterial liquid was 5×10 5 CFU/mL by a 2-fold dilution method. The 96-well plate is placed in a biochemical incubator for culturing for 24 hours, the minimum dilution concentration of the drug without obvious bacterial growth is MIC (mg/mL) of the drug, and the minimum dilution concentration of the drug with aseptic growth after 24 hours of drip plate culturing is MBC (mg/mL) of the drug.
The results are shown in Table 1, and compared with comparative examples 1-2, examples 1-3 have better antibacterial activity on staphylococcus aureus, salmonella and escherichia coli, which shows that the proportion of the traditional Chinese medicine composition in the range of the embodiment of the invention has more advantages in the aspect of in vitro bacteriostasis; as shown in the comparison result with comparative example 3, the callicarpa nudiflora can greatly improve the antibacterial activity on salmonella and escherichia coli in the formula of the invention; in comparison with comparative examples 4 to 5, it was found that pericarpium Granati and radix Sanguisorbae are more suitable than Galla chinensis to improve the antibacterial activity of the composition; in comparison with comparative example 6, it was found that the traditional Chinese medicines of honeysuckle and weeping forsythiae, which have the same effects of clearing heat and detoxicating as the coptis and the baical skullcap root, are used for replacing the coptis and the baical skullcap root, wherein the in vitro antibacterial activity of the medicine composition is weaker; in comparison with comparative example 7, the antibacterial activity of group Fang Zhongbai pulsatilla is lower than that of the combination of radix scutellariae and garden burnet, which shows that under the same quality, the antibacterial effect of radix scutellariae and garden burnet in vitro is better after the combination than that of the pulsatilla alone; in comparison with comparative example 8, it was found that the use of papain is more conducive to dissolution of bacteriostatic substances than the complex enzymatic hydrolysis of amylase with cellulase and pectinase; in comparison with comparative example 9, it was found that an enzymolysis ratio outside the technical range of the invention can weaken the bacteriostatic activity of the formulation and reflect the important role of papain in the composite enzymolysis extraction process.
Table 1 comparison of in vitro bacteriostatic activity of the Chinese medicinal compositions of examples 1 to 3 and comparative examples 1 to 9
2. Example 1 in vitro bacteriostatic efficacy comparison assessment with prior art patents and classical formulations
Evaluation example 1 was conducted in the same manner as in the micro broth dilution method in the above "1. Evaluation of in vitro bacteriostatic activity", with patent document CN103830400a (coptis chinensis 1kg, phellodendron bark 1kg, scutellaria baicalensis 1kg, andrographis paniculata 1kg, bai Weng 2kg, atractylodes macrocephala 2kg, white peony root 2kg, pericarpium Granati 2kg, dandelion 2kg, licorice 2kg, sanguisorba officinalis 2kg, magnolia officinalis 0.5 kg, halloysite 0.5 kg, kochia scoparia 0.5 kg, isatis root 2.5 kg, dyer woad leaf 2.5 kg, dried orange peel 2.5 kg, reference example 1 preparation method) and CN103006916A (white peony root 1g, white atractylodes rhizome 1g, pulsatilla root 1g, atractylodes rhizome 1g, dried orange peel 2 g, dyers woad leaf 1g, phellodendron bark 1g, weeping forsythiae capsule 0.75 g, dandelion 3g, ash bark 1.5 g, garden burnet 5g, pomegranate rind 1g, poria cocos 1g, reference example 1 preparation method), CN g a (polygonum orientalis 350 g, spider aroma 200 g, oleaster leaf 200 g, bistort 150 g, cactus 100g, reference example 1 preparation method), and comparing with classical anti-piglet diarrhea oral liquid-pulsatilla root oral liquid (pulsatilla root 300 g, coptis root 150 2, ash bark 300 g, phellodendron bark 225 g); in vitro bacteriostatic activity MIC (mg/mL) and MBC (mg/mL) were obtained from Guangdong Dahua farm animal healthcare Co., ltd.) and poplar flower oral liquid (poplar flower extraction mixture; from Huishuang Biotech Co., ltd.).
TABLE 2
As shown in Table 2, the in vitro antibacterial activity of the Chinese medicinal composition in the invention for staphylococcus aureus, salmonella and escherichia coli is higher than that of the Chinese medicinal composition in the patent document CN103830400A, CN103006916A, CN108938832A, and is also higher than that of the Chinese pulsatilla oral liquid and the poplar flower oral liquid, and although the Chinese medicinal compound in the patent document CN103830400A is similar to the Chinese medicinal composition in the invention, the Chinese medicinal composition has obvious in vitro antibacterial activity advantage.
3. Evaluation of clinical preventive Effect
Location: heshan Hongfa farm and grazing Co.Ltd
Test object: 100 healthy primary piglets of 3 days old
Test grouping: 1) Blank 10 heads (without any drug); test 1 group 10 heads (using example 1 formulation of the herbal composition); test 2 to 4 groups of 10 heads each (using the traditional Chinese medicine compositions of comparative examples 2,3, 5; each of the test 5 to 6 groups was 10 (each using a preparation method of the Chinese medicinal composition of patent document CN 103830400A: 1kg of coptis chinensis, 1kg of phellodendron bark, 1kg of scutellaria baicalensis, 1kg of andrographis paniculata, bai Weng kg of dyers woad leaf, 2kg of atractylodes macrocephala koidz, 2kg of white paeony root, 2kg of pericarpium Granati, 2kg of dandelion, 2kg of liquorice, 2kg of sanguisorba officinalis, 0.5kg of magnolia officinalis, 0.5kg of Halloysitum rubrum, 0.5kg of lithocarpus, 2.5kg of radix isatidis, 2.5kg of dyers woad leaf, 2.5kg of dried orange peel, reference example 1; a preparation method of the Chinese medicinal composition of patent document CN 103006916A: 1g of white paeony root, 1g of atractylodes macrocephala koidz, 1g of pulsatilla chinensis, 1g of rhizoma atractylodis, 2g of dried orange peel, 1g of dyers woad leaf, 1g of phellodendron bark, 0.75g of fructus forsythiae, 3g of dandelion, 1.5g of cortex fraxini, 5g of garden burnet, 1g of pericarpium Granati, 1g of poria cocos, and reference to the preparation method of comparative example 7); 10 groups of experiments 7 to 8 (commercial control group: pulsatilla root oral liquid and poplar flower oral liquid); test 9 groups 10 heads (model control group).
The test method comprises the following steps: carrying out oral liquid infusion administration on the tested piglets in the test 1-9 groups, wherein the drug concentration in the test 1-6 groups is 0.5g/mL, the administration dosage is 0.5mL/kg body weight, the drug concentration in the test 7-8 groups is 1g/mL, and the administration dosage is 0.25mL/kg body weight; after the administration, 2mL of the escherichia coli K88 bacterial liquid with the concentration of 10 8 CFU/mL is filled into the tested pigs of the test 1-9 groups for 3 days, the continuous observation is carried out for 7 days, and indexes such as mental state, diarrhea rate, production performance and the like of the tested pigs are recorded, so that the effect advantage of the traditional Chinese medicine composition of the formula of the embodiment 1 for clinically preventing the yellow diarrhea of piglets is verified.
Results:
(1) Mental state: the mental state of the tested pigs of 1-8 groups before and after administration is free from abnormality, and the test pigs are particularly sensitive in response and active in milk taking.
(2) Diarrhea rate: the diarrhea rate of piglets is observed by filling escherichia coli after administration of the pigs, the occurrence judgment standard is shown in figure 1, the statistics of diarrhea of pigs after administration of the medicines are shown in table 3, the pigs of each group have no diarrhea before administration, and the pigs of each group have partial diarrhea after administration, so that the factors are small and the individual difference is considered to be caused. In 7 days after the administration of Escherichia coli, the diarrhea rate of pigs in test 9 groups (model group) is found to be 90%, compared with the diarrhea rate of pigs in test 1-8 groups which are lower than that of pigs in test 9 groups, wherein the diarrhea rate of pigs in test 1 group (example 1) is the lowest, the prevention effect is the best, and the clinical prevention effect of the formula ratio of the invention is proved to be better than that of patent document CN103830400A again.
Table 3 statistics of diarrhea rate of different test nodes in the herd
| Group of | Pre-dose (%) | Post-administration (%) | After molding (%) |
| Blank group | 0 | 0 | 0 |
| Test 1 group | 0 | 0 | 10 |
| Test 2 groups | 0 | 10 | 40 |
| Test 3 groups | 0 | 10 | 50 |
| Test 4 groups | 0 | 10 | 50 |
| Test 5 groups | 0 | 0 | 40 |
| Test 6 groups | 0 | 0 | 50 |
| Test 7 groups | 0 | 10 | 70 |
| Test 8 groups | 0 | 0 | 40 |
| Test 9 groups | 0 | 10 | 90 |
(3) The production performance is as follows: the related indexes are shown in table 4, after diarrhea occurs in most tested pig groups in the test group, compared with the blank group, growth is delayed, but the daily gain of the test 1 group and the daily gain of the test 8 group are not obviously different (p is more than 0.05), which indicates that the traditional Chinese medicine composition and the poplar flower oral liquid in the embodiment 1 can effectively prevent adverse effects caused by escherichia coli infection; the daily gain of each test group is obviously lower than that of a blank group and a test 1 group (p is less than 0.05), which indicates that the traditional Chinese medicine preparation in the example 1 has better clinical prevention effect compared with other comparative examples, the existing patent documents and the pulsatilla root oral liquid.
Table 4 weighing statistics of the pigs tested
Note that: the same letter represents no significant difference between groups (p > 0.05) and the different letters represent significant differences between groups (p < 0.05).
4. Evaluation of clinical treatment Effect
Location: heshan Hongfa farm and grazing Co.Ltd
Test object: 200 healthy piglets of 3-5 days old.
Test grouping: 1) Blank 20 heads (without any drug); test 1 group 20 heads (using example 1 formulation of the herbal composition); 20 heads of each of the test 2-4 groups (using the traditional Chinese medicine compositions of the compositions of comparative examples 2,3 and 5); each group of 20 groups (20 groups of experiments respectively using the Chinese medicinal composition in patent document CN 103830400A: 1kg of coptis chinensis, 1kg of phellodendron bark, 1kg of scutellaria baicalensis, 1kg of common andrographis herb, bai Weng kg of Chinese angelica, 2kg of bighead atractylodes rhizome, 2kg of white paeony root, 2kg of pericarpium Granati, 2kg of dandelion, 2kg of liquorice, 2kg of garden burnet root, 0.5kg of magnolia officinalis, 0.5kg of red halloysite, 0.5kg of lithocarpus fruit, 2.5kg of radix isatidis, 2.5kg of dyers woad leaf, 2.5kg of dried orange peel, reference example 1 preparation method, and the Chinese medicinal composition in patent document CN 103006916A: 1g of white paeony root, 1g of bighead atractylodes rhizome, 1g of Chinese pulsatilla root, 1g of rhizoma atractylodis, 2g of dried orange peel, 1g of dyers woad leaf, 1g of phellodendron bark, 0.75g of fructus forsythiae, 3g of dandelion, 1.5g of cortex fraxini, 5g of garden burnet root, 1g of pericarpium Granati, 1g of poria cocos, and reference to the preparation method of comparative example 7); 20 groups of experiments 7 to 8 (commercial control group: pulsatilla root oral liquid and poplar flower oral liquid); test 9 group 20 heads (model control group).
The test method comprises the following steps: 2mL of coliform K88 bacteria solution with the concentration of 10 8 CFU/mL is filled into the test 1-9 groups of piglets, the test 1-6 groups of piglets are subjected to mold making, and the test 1-6 groups of piglets begin to be filled after yellow thin feces are discharged, wherein the concentration of the test 1-8 groups of piglets is 0.5g/mL, and the concentration of the test 7-8 groups of piglets is 1g/mL; the doses of 1-6 groups were 1mL/kg body weight, the doses of 7-8 groups were 0.5mL/kg body weight, 1 time daily, and after 3 days of continuous administration, the diarrhea rate, cure rate, daily gain, and blood biochemical index were recorded, the clinical treatment effect of comparative example 1 and other comparative examples, published patents, and classical formulations were evaluated, the cure judgment criteria was that anus was relatively dry, and no yellow thin feces were squeezed to spill (FIG. 2).
As shown in Table 5, the K88 strain has a stable molding diarrhea rate of more than 95% (test 1-9 groups), and by administration observation, all test groups can remarkably improve the cure rate of diarrhea piglets compared with the model group (test 9 group), and compared with the test 1 group, the cure rate is highest, so that the diarrhea piglet has an optimal therapeutic preparation. In statistics of daily gain, it was found that the model group significantly decreased compared to the blank group, indicating that the E.coli infection significantly retarded the growth of piglets, whereas the drug-treated group (test 1 group to 8 group) would alleviate the growth retardation of piglets to some extent, especially test 1 group, the daily gain was significantly higher than the model group (p < 0.05) and there was no significant difference (p > 0.05) compared to the blank group.
Table 5 piglet yellow dysentery modeling and treatment effect statistics
Note that: the same letter represents no significant difference between groups (p > 0.05) and the different letters represent significant differences between groups (p < 0.05).
Further, the blood routine and biochemical detection is carried out on the tested pig groups in the blank group, the test 1 group and the model group, the results are shown in the table 6, the blood routine data and the serum biochemical parameters of the tested pig groups before and after the administration are in the normal range, and compared with the blank group and the model group, no obvious difference exists, so that the traditional Chinese medicine composition has no obvious influence on the blood of piglets.
TABLE 6 blood routine and Biochemical detection results
5. Evaluation of clinical treatment Effect (II)
Location: guangdong province Maoque market certain pig farm
Test object: 150 primary piglets (5 days old) are discharged with yellow dilute manure and mental depression, and are identified by epidemiological investigation, symptom analysis and laboratory separation and identification, and are identified as enterotoxigenic escherichia coli infection and piglet yellow dysentery.
Test grouping: 1) Blank 30 heads (without any drug); test 1 group 30 heads (using example 1 formulation of the Chinese medicinal composition); 30 heads of test group 2 (using the Chinese medicinal composition of patent document CN 103830400A: coptis root 1kg, phellodendron bark 1kg, baikal skullcap root 1kg, andrographis paniculata 1kg, bai Weng kg, atractylodes macrocephala 2kg, white peony root 2kg, pomegranate rind 2kg, dandelion 2kg, licorice 2kg, garden burnet root 2kg, magnolia bark 0.5kg, red halloysite 0.5kg, lithocarpus 0.5kg, isatis root 2.5kg, dyers woad leaf 2.5kg, dried orange peel 2.5kg, reference to the preparation method of example 1); 30 groups of experiments 3 to 4 (commercial control group: pulsatilla root oral liquid and poplar flower oral liquid).
The test method comprises the following steps: the tested piglets in the test 1-4 groups are administrated by drenching, the drug concentration in the test 1-2 groups is 0.5g/mL, the administration dosage is 1mL/kg body weight, the drug concentration in the test 3-4 groups is 1g/mL, and the administration dosage is 0.5mL/kg body weight; the administration is continued for 3 days, and the observation is carried out for 7 days, the diarrhea rate, the cure rate and the daily gain of the tested pigs are recorded, and the advantage of the clinical treatment effect of the traditional Chinese medicine preparation of the formula 1 in the embodiment on the yellow diarrhea of piglets is checked.
Results: as shown in Table 7, the daily gain of the blank group is obviously lower than that of other test groups, the growth speed of the piglets is obviously influenced after the piglets are subjected to yellow dysentery, the cure rate of the test group is obviously higher than that of the control group (p < 0.05) after the piglets are subjected to medication treatment, the daily gain of the test group 1 is highest, and the effect of the invention in the yellow dysentery of the piglets is better compared with that of the piglets in other patents and classical formulas.
Table 7 piglet yellow dysentery modeling and treatment effect statistics
Note that: the same letter represents no significant difference between groups (p > 0.05) and the different letters represent significant differences between groups (p < 0.05).
6. Evaluation of clinical treatment Effect (III)
Location: pig farm in Yangjiang city of Guangdong province
Test object: 100 weaned pigs (27 days old) are treated by white dilute manure discharge, mental depression, epidemiological investigation, symptom analysis and laboratory separation and identification, and are determined to be enterotoxigenic escherichia coli infection and white diarrhea of the piglets.
Test grouping: 1) Blank 20 heads (without any drug); test 1 group 20 heads (using example 1 formulation of the herbal composition); group 2, 20 heads (using the Chinese medicinal composition of patent document CN 103830400A: coptis root 1kg, phellodendron bark 1kg, baikal skullcap root 1kg, andrographis paniculata 1kg, bai Weng kg, atractylodes macrocephala 2kg, white peony root 2kg, pomegranate rind 2kg, dandelion 2kg, licorice root 2kg, garden burnet root 2kg, magnolia bark 0.5kg, red halloysite 0.5kg, lithocarpus 0.5kg, isatis root 2.5kg, dyers woad leaf 2.5kg, dried orange peel 2.5kg, reference example 1 preparation method); 20 groups of experiments 3 to 4 (commercial control group: pulsatilla root oral liquid and poplar flower oral liquid).
The test method comprises the following steps: the tested piglets in the test 1-4 groups are administrated by drenching, the drug concentration in the test 1-2 groups is 0.5g/mL, the administration dosage is 1mL/kg body weight, the drug concentration in the test 3-4 groups is 1g/mL, and the administration dosage is 0.5mL/kg body weight; the administration is continued for 3 days, and the observation is carried out for 7 days, the diarrhea rate, the cure rate and the daily gain of the tested pigs are recorded, and the advantage of the clinical treatment effect of the traditional Chinese medicine preparation of the formula 1 in the embodiment on the white diarrhea of piglets is checked.
Results: as shown in table 8, the daily gain of the blank group is significantly lower than that of other test groups, which indicates that the piglets need to be intervened in time after white diarrhea, in the test, the cure rate of the test group is significantly higher than that of the control group (p < 0.05), wherein the cure rate of the test group 1 and the daily gain of the piglets are the highest, which indicates that compared with other published patents and classical formulas, the effect of the embodiment 1 of the invention on the white diarrhea of weaned piglets is better.
Table 8 statistics of yellow diarrhea modeling and treatment effects of piglets
Note that: the same letter represents no significant difference between groups (p > 0.05) and the different letters represent significant differences between groups (p < 0.05).
7. Comprehensive influence of different preparation methods on curative effect of traditional Chinese medicine composition
1) Evaluation of the enhancing effect of the preparation method of the Chinese medicinal composition on the extraction of the effective ingredients of the medicinal materials
In order to verify the advantages of the preparation method of the traditional Chinese medicine composition compared with the conventional preparation method, corresponding orthogonal tests are designed for comprehensive evaluation. Taking example 1 as an example, firstly, smashing medicinal materials involved in the formulas of callicarpa nudiflora, pericarpium Granati, coptis chinensis and the like, respectively weighing callicarpa nudiflora 12 g, coptis chinensis 10 g, scutellaria baicalensis 15 g, pericarpium Granati 15 g, sanguisorba 15 g and liquorice 10 g, taking 1 part, weighing 7 parts in total, and the preparation method comprises 3 extraction modes, and evaluating the optimal extraction method by adopting an orthogonal test method, wherein the ultrasonic extraction temperature is 55 ℃, the power is 180W, and the extraction time is 40 min; the compound enzymolysis extraction involves cellulase (10 ten thousand U/g): pectase (9U/g): xylose protease (10 ten thousand U/g) mass ratio=1:1.5:3, the addition ratio is 4% of the medicinal material mass; the conventional water extraction adopts a boiling extraction method (the baical skullcap root is added after being boiled), the ultrasonic extraction, the composite enzymolysis extraction and the conventional water extraction are sequentially carried out according to the sequence of the ultrasonic extraction, the composite enzymolysis extraction and the conventional water extraction, the test is divided into 7 different preparation methods, and the preparation samples 1-7 are prepared, and the specific grouping is shown in table 9.
Table 9 preparation method investigation grouping
Collecting the Chinese medicinal extract, and selectively comparing the contents of baicalin and berberine hydrochloride in 7 extraction methods according to high performance liquid chromatography, wherein the content calculation formula is as follows (in the calculation process, the content of the reference substance is higher (approaching 100%), so that the reference substance is regarded as 1):
The results of the detection of baicalin and berberine hydrochloride content in samples 1 to 7 are shown in tables 10 to 13 and figures 3 to 4.
Table 10 Peak results (baicalin)
Table 11 Peak results (berberine hydrochloride)
Table 12 baicalin content measurement (HPLC method) results
Note that: a represents that the baicalin content in the sample has a significant difference compared with the sample 1, and p is less than 0.05.
TABLE 13 determination of berberine hydrochloride content (HPLC method) results
Note that: a represents that the baicalin content in the sample has a significant difference compared with the sample 1, and p is less than 0.05.
Results: the content of baicalin and berberine hydrochloride in the traditional Chinese medicine preparation prepared by single ultrasonic extraction or compound enzymolysis extraction is obviously lower than that of the traditional Chinese medicine preparation prepared by single water extraction (p < 0.05), but before the water extraction, the content of baicalin and berberine hydrochloride in the traditional Chinese medicine preparation prepared by single water extraction can be obviously improved by ultrasonic auxiliary compound enzymolysis extraction of the medicinal powder (p < 0.05).
2) Evaluating the in vitro bacteriostatic effect of samples 1-7
The in vitro bacteriostatic effect of samples 1 to 7 was further evaluated by the oxford cup method, and the specific steps were as follows:
In an ultra clean bench, pour about 10mL of the MH agar culture medium sterilized under pressure into each sterilization culture dish to pave the bottom, after the bottom agar is cooled and solidified, place 4 sterilized oxford cups on the agar, and the interval should be equal. 1mL of the indicator bacterium liquid with the absorbance value of OD 600 being 0.1 is sucked and added into 100mL of MH agar medium with the constant temperature of 50+/-5 ℃ and gently shaken to avoid foaming, and the concentration of the indicator bacterium in the medium is 10 6 CFU/mL. The MH agar medium with the bacterial liquid is poured into MH agar plates with oxford cups, 10mL of each plate is poured, and the layers are uniformly paved and cooled and solidified. 200. Mu.L of the drug to be tested (concentration of 0.5 mg/mL) was pipetted into the corresponding oxford cup while sterile water was set up as a blank and marked on the petri dish. And (3) standing the flat plate added with the medicine to be tested in a low-temperature environment at 4-10 ℃ for pre-permeation for 4-6 hours, and waiting for the liquid in the oxford cup to permeate into the agar. The permeated culture dish was transferred to a 37℃incubator for cultivation. Observing the growth condition and the antibacterial effect of thalli in the next 6-16 h; after the inhibition zone is found, the inhibition zone is measured and recorded by using a vernier caliper.
The test results are shown in Table 14, samples 2-4 have no obvious enhancement (p > 0.05) of bacteriostatic ability compared with sample 1, which indicates that the use of ultrasonic extraction or compound enzymolysis extraction does not enhance the bacteriostatic activity of the traditional Chinese medicine preparation compared with the traditional water extraction; the in-vitro antibacterial activity (p < 0.05) of the traditional Chinese medicine preparation can be obviously increased by ultrasonic-assisted compound enzymolysis extraction before water extraction, which proves that the extraction method can promote the antibacterial substances in the traditional Chinese medicine to be dissolved out to a greater extent.
Table 14 antibacterial results of Chinese medicinal preparation
Note that: the same letter represents no significant difference between groups (p > 0.05) and the different letters represent significant differences between groups (p < 0.05).
3) Clinical test evaluation samples 1 to 7 have different therapeutic effects on piglet yellow scour
The yellow and white thin feces appear in 150 piglets of 300 piglets of Heshan Hongfa farm and animal husbandry limited company of Guangdong province, and the yellow and white diarrhea of the piglets is confirmed by clinical symptom analysis and laboratory diagnosis, and the effect difference of clinically treating the yellow diarrhea of the piglets is evaluated by drenching samples 1-7.
Test grouping: control group 10 heads (no drug); test 1 to 7 groups of 20 heads (drenched samples 1 to 7).
The test method comprises the following steps: and (3) carrying out oral liquid infusion administration on the tested piglets of the test groups 1-7, wherein the drug concentration is 0.5g/mL, the administration dosage is 1mL/kg body weight, the administration is continuously carried out for 3 days, the observation is carried out for 7 days, the diarrhea rate, the cure rate and the daily gain of the tested piglets are recorded, and the effect advantage of the traditional Chinese medicine preparation of the formula of the embodiment 1 for clinically treating the yellow-white diarrhea of the piglets is checked.
As shown in Table 15, the daily gain of the blank group is significantly lower than that of other test groups, which indicates that the piglets need to be intervened in time after yellow and white dysentery, in the test, the cure rate of the test 1-7 groups is significantly higher than that of the control group (p < 0.05), wherein the cure rate and the daily gain of the test 7 groups are highest, which indicates that compared with other samples, the sample 7 of the invention has better effect of clinically treating the yellow and white dysentery of the piglets, and the application of the ultrasonic-assisted composite enzymolysis extraction technology can promote the clinical curative effect of the traditional Chinese medicine preparation.
Table 15 piglet yellow dysentery modeling and treatment effect statistics
Note that: the same letter represents no significant difference between groups (p > 0.05) and the different letters represent significant differences between groups (p < 0.05).
To sum up: the results of experiments 1-2 show that compared with other orthogonal groups, the traditional Chinese medicine composition prepared by ultrasonic-assisted composite enzymolysis extraction can remarkably improve the content of active ingredients in the traditional Chinese medicine composition and the in-vitro antibacterial activity of the traditional Chinese medicine composition; the test 3 shows that the higher content of the active ingredients and the in-vitro antibacterial activity can ensure better clinical treatment effect, and the higher clinical cure rate and daily gain are particularly realized, thereby having important significance for promoting the normal growth level of piglets suffering from yellow-white dysentery to be recovered. Therefore, the preparation method of the traditional Chinese medicine composition selects ultrasonic-assisted compound enzymolysis extraction, so that the extraction efficiency and clinical curative effect of the traditional Chinese medicine can be remarkably improved.
While the embodiments of the present invention have been described in detail, the present invention is not limited to the above embodiments, and various changes can be made without departing from the spirit of the present invention within the knowledge of those skilled in the art. Furthermore, embodiments of the invention and features of the embodiments may be combined with each other without conflict.
Claims (6)
1. A traditional Chinese medicine composition for preventing and treating swine yellow and white dysentery is characterized by being prepared from the following raw materials in parts by weight: 10-15 parts of callicarpa nudiflora, 8-15 parts of coptis chinensis, 12-20 parts of radix scutellariae, 10-18 parts of pericarpium granati, 10-20 parts of radix sanguisorbae and 8-12 parts of liquorice; the traditional Chinese medicine composition is prepared by a preparation method comprising the following steps:
Taking the powder of beautyberry, baical skullcap root, golden thread, pomegranate rind, garden burnet root and liquoric root according to the proportion,
S1: mixing powder of Callicarpa nudiflora, coptidis rhizoma, pericarpium Granati, radix Sangusorbae and Glycyrrhrizae radix, adding water, ultrasonic extracting, and performing enzymolysis with complex enzyme to obtain extractive solution A and residue B;
S2: extracting Scutellariae radix powder with water to obtain extractive solution C and residue D;
s3: mixing the residue B and the residue D, extracting with water to obtain extractive solution E;
s4: mixing the extract A, the extract C and the extract E, and concentrating to obtain the traditional Chinese medicine composition;
The complex enzyme in S1 is cellulase, pectase and papain, and the mass ratio of the cellulase to the pectase to the papain is (1-2): (1-2): (3-4); the enzyme activity of each enzyme in the complex enzyme is 10-15 ten thousand U/g;
The enzymolysis conditions of the complex enzyme in S1 are as follows: the pH value is 4.5-5.0, the temperature is 50-60 ℃ and the time is 1-3 h.
2. The traditional Chinese medicine composition for preventing and treating swine yellow and white dysentery according to claim 1, wherein the traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 11-13 parts of callicarpa nudiflora, 9-11 parts of coptis chinensis, 13-15 parts of radix scutellariae, 12-16 parts of pericarpium granati, 14-16 parts of garden burnet and 9-11 parts of liquorice.
3. The method for preparing the traditional Chinese medicine composition for preventing and treating swine yellow and white dysentery as claimed in claim 1 or 2, which is characterized by comprising the following steps:
Taking the powder of beautyberry, baical skullcap root, golden thread, pomegranate rind, garden burnet root and liquoric root according to the proportion,
S1: mixing powder of Callicarpa nudiflora, coptidis rhizoma, pericarpium Granati, radix Sangusorbae and Glycyrrhrizae radix, adding water, ultrasonic extracting, and performing enzymolysis with complex enzyme to obtain extractive solution A and residue B;
S2: extracting Scutellariae radix powder with water to obtain extractive solution C and residue D;
s3: mixing the residue B and the residue D, extracting with water to obtain extractive solution E;
s4: mixing the extract A, the extract C and the extract E, and concentrating to obtain the traditional Chinese medicine composition;
The temperature of ultrasonic extraction in the step S1 is 50-60 ℃, the power of ultrasonic extraction is 100-180W, and the time is 30-40 min;
The adding amount of the complex enzyme in the S1 is 3% -5% of the mass of the extracting solution A;
The complex enzyme in S1 is cellulase, pectase and papain, and the mass ratio of the cellulase to the pectase to the papain is (1-2): (1-2): (3-4); the enzyme activity of each enzyme in the complex enzyme is 10-15 ten thousand U/g; the enzymolysis conditions of the complex enzyme in S1 are as follows: the pH value is 4.5-5.0, the temperature is 50-60 ℃ and the time is 1-3 h.
4. A medicament for preventing and treating swine yellow and white dysentery, which is characterized in that active ingredients in the medicament comprise the traditional Chinese medicine composition of claim 1 or 2.
5. The drug for preventing and treating swine yellow and white dysentery of claim 4, further comprising pharmaceutically acceptable excipients.
6. Use of a Chinese medicinal composition according to claim 1 or 2 for the preparation of a medicament for the prophylaxis and/or treatment of at least one of (a) to (c):
(a) Yellow dysentery of pigs;
(b) White diarrhea of pigs;
(c) Bacterial infection in pigs.
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